Simultaneous determination of the flavonoids robinin and kaempferol in human breast cancer cells by liquid chromatography-tandem mass spectrometry

J Pharm Biomed Anal. 2011 Apr 28;55(1):109-13. doi: 10.1016/j.jpba.2010.12.021. Epub 2010 Dec 23.


An accurate, precise and sensitive method was developed and validated for the simultaneous quantification of the flavonoid glycoside robinin, and its algycone kaempferol in human breast cancer MCF-7 cells. The application of liquid chromatography-tandem mass spectrometry (LC/MS/MS) with a TurboIonspray interface in negative mode under multiple reactions monitoring was investigated. Chromatographic separation was achieved on a C(18) column using a mobile phase consisting of (A) water with 0.025% formic acid and 1mM ammonium formate and (B) acetonitrile with 0.025% formic acid. Rutin was used as the internal standard for robinin and fisetin as the internal standard for kaempferol. The assay had a limit of detection of 0.1ng/ml for both compounds when present in cell lysate. The calibration curves were linear from 1 to 250ng/ml (r>0.999) for each compound. The intra- and inter-day coefficients of variation were less than 10% and intra- and inter-day accuracies were within 11%. This assay was successfully applied in a robinin cellular uptake study to determine the intracellular concentrations of robinin in MCF-7 cells.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Anti-Inflammatory Agents, Non-Steroidal / metabolism
  • Anti-Inflammatory Agents, Non-Steroidal / pharmacokinetics
  • Biological Transport
  • Breast Neoplasms / metabolism*
  • Calibration
  • Cell Line, Tumor
  • Chromatography, High Pressure Liquid
  • Drug Stability
  • Female
  • Flavonoids / metabolism*
  • Flavonoids / pharmacokinetics
  • Humans
  • Kaempferols / metabolism*
  • Limit of Detection
  • Microchemistry / methods*
  • Osmolar Concentration
  • Reproducibility of Results
  • Spectrometry, Mass, Electrospray Ionization
  • Tandem Mass Spectrometry


  • Anti-Inflammatory Agents, Non-Steroidal
  • Flavonoids
  • Kaempferols
  • kaempferol
  • robinin